Sequence microheterogeneity of parvalbumin pI 5.0 of pike: a mass spectrometric study.
Parvalbumin (PA) is a muscle and neuronal calcium-binding protein, the major fish and frog allergen. Its characteristic feature is the presence of multiple isoforms with significantly different amino acid sequences. Here we show that the major isoform of northern pike muscle PA (pI 5.0, alpha-PA) exhibits microheterogeneity of amino acid sequence. ESI Q-TOF mass-spectrometry (MS) analysis of alpha-PA sample showed the presence of two components with mass difference of 71 Da. Analysis of tryptic and endoproteinase Asp-N digests of alpha-PA by MALDI-TOF MS revealed peptides, corresponding to two different amino acid sequences. The sequence differences between variant proteins are limited to AB-domain and include substitutions K27A and L31K, and an extra Leu residue between K11 and K12. Since the affected residues comprise a cluster on the surface of PA, an involvement of the identified region into target recognition is suggested. The substitutions at positions 27 and 31 are located in the region of previously identified epitopes of parvalbumin relevant for PA-specific IgE and IgG binding, which suggests different immunoactivities of the variants. The found microheterogeneity of PA is suggested to be of importance for physiological adaptation of the propulsive musculature to developmental and/or environmental requirements and may contribute to PA allergenicity.
Permyakov
Sergei E
SE
Institute for Biological Instrumentation of the Russian Academy of Sciences, Pushchino, Moscow Region 142290, Russia. pse101@itaec.ru
Karnoup
Anton S
AS
Bakunts
Anush G
AG
Permyakov
Eugene A
EA
D000485:Allergens / Q000276:immunology / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D017859:Esocidae; D007526:Isoelectric Point; D007930:Leucine / Q000737:chemistry; D008666:Metalloendopeptidases / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010320:Parvalbumins / Q000737:chemistry* / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry; D020033:Protein Isoforms / Q000737:chemistry / Q000378:metabolism; D020539:Sequence Analysis, Protein; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors; D014357:Trypsin / Q000378:metabolism
D000485:Allergens; D010320:Parvalbumins; D010446:Peptide Fragments; D020033:Protein Isoforms; D014357:Trypsin; D008666:Metalloendopeptidases; C058921:endoproteinase Asp-N; D007930:Leucine